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• W1992299998 endingPage "20472" @default.
• W1992299998 startingPage "20465" @default.
• W1992299998 abstract "Erythropoietin (Epo) activates a voltage-independent Ca²⁺ channel that is dependent on tyrosine phosphorylation. To identify the domain(s) of the Epo receptor (Epo-R) required for Epo-induced Ca²⁺ influx, Chinese hamster ovary (CHO) cells were transfected with wild-type or mutant Epo receptors subcloned into pTracer-cytomegalovirus vector. This vector contains an SV40 early promoter, which drives expression of the green fluorescent protein (GFP) gene, and a cytomegalovirus immediate-early promoter driving expression of the Epo-R. Successful transfection was verified in single cells by detection of GFP, and intracellular Ca²⁺ ([Ca]_i) changes were simultaneously monitored with rhod-2. Transfection of CHO cells with pTracer encoding wild-type Epo-R, but not pTracer alone, resulted in an Epo-induced [Ca]_i increase that was abolished in cells transfected with Epo-R F8 (all eight cytoplasmic tyrosines substituted). Transfection with carboxyl-terminal deletion mutants indicated that removal of the terminal four tyrosine phosphorylation sites, but not the tyrosine at position 479, abolished Epo-induced [Ca]_i increase, suggesting that tyrosines at positions 443, 460, and/or 464 are important. In CHO cells transfected with mutant Epo-R in which phenylalanine was substituted for individual tyrosines, a significant increase in [Ca]_i was observed with mutants Epo-R Y443F and Epo-R Y464F. The rise in [Ca]_i was abolished in cells transfected with Epo-R Y460F. Results were confirmed with CHO cells transfected with plasmids expressing Epo-R mutants in which individual tyrosines were added back to Epo-R F8 and in stably transfected Ba/F3 cells. These results demonstrate a critical role for the Epo-R cytoplasmic tyrosine 460 in Epo-stimulated Ca²⁺ influx." @default.
• W1992299998 created "2016-06-24" @default.
• W1992299998 creator A5000828286 @default.
• W1992299998 creator A5021303868 @default.
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• W1992299998 date "1999-07-01" @default.
• W1992299998 modified "2023-10-12" @default.
• W1992299998 title "Identification of the Erythropoietin Receptor Domain Required for Calcium Channel Activation" @default.
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• W1992299998 doi "https://doi.org/10.1074/jbc.274.29.20465" @default.
• W1992299998 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/10400674" @default.
• W1992299998 hasPublicationYear "1999" @default.
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